Small Mucous Granule Research Articles

Morphology of adhesive organ of the snow trout Schizothorax richardsonii (Gray, 1832)

This paper describes the morphology and fine structure of adhesive organ (AO) of the snow trout Schizothorax richardsonii (Gray, 1832). The species lives in the sub‐Himalayan streams and rivers, where they adhere to submerged rocks and stones with its AO located beneath the mouth opening. The AO consists of a crescent callus part and a modified labial fold, bearing numerous tubercles. In both parts, the epidermis is exceptionally thick (150–170 µm) and contains keratinocytes rich in bundles of tonofilaments; elsewhere, the epidermis contains single, disperse filaments, mucous cells and other usual cells seen in teleosts. Unlike epidermis elsewhere, the outer 4–5 row cells of the AO epidermis undergo keratinisation and are cytokeratin positive. The individual keratinised cell layers, which are degenerated electron‐dense sheets, lack nuclei and cytoplasmic organelles, but possess filaments embedded in a dense cytoplasmic matrix and few, large mucous‐like granules. Beneath the keratinised layers, keratinocytes are compactly aligned into 20–25 rows. They possess abundant thick bundles of clumped tonofilaments over the entire cytoplasm and peripherally, small mucous granules. Towards the basal epidermis, the cells are elongated and show major cytoplasmic organelles, few tonofilament bundles, but no mucous granules. It is evident that the keratinocytes of the mid‐epidermis are differentiated for the production of tonofilaments. The outer keratinised part of the AO epidermis provides structural support and protection (from damage) during adhesion. It appears that the callus part acts by creating vacuum first, while the tuberculated part of the AO epidermis assists in adhesion to the rocky substratum.

Structure of adhesive organ of the mountain‐stream catfish, Pseudocheneis sulcatus (Teleostei: Sisoridae)

AbstractThe structure and ultrastructure of the adhesive organ (AO) in the catfish, Pseudocheneis sulcatus (Sisoridae), an inhabitant of the sub‐Himalayan streams of India, is described. The surface of the AO is thrown into folds, the ridges of which bear curved spines. The AO epidermis consists of 10–12 tiers of filament‐rich cells, of which the outer tier cells project spines lined with a thick plasma membrane and bear bundles of tonofilaments (TF). Their cytoplasm contains TF and large mucus‐like granules, but no obvious organelles. A second tier of living cells with spines is present beneath the outer tier and seems to replace the latter when its spines are damaged or shed. The outer tier cells react positively with antibody to cytokeratin. Actin labelling is clearly absent from the outer tier, indicating that keratinization of the outer tier occurs in the absence of actin filaments. In the cells of the third to fifth tiers, the cytoplasm possesses abundant small mucous granules (0.1–0.3 µm), and fewer TF compared to the cytoplasm in the spines. The cells of the innermost tiers and the basal layer possess few TF bundles, but no mucous granules. The potential of AO filament cells to produce both mucous granules and keratin filaments is noteworthy. The observations provide evidence that specific regions of fish epidermis can actually undergo a true process of keratinization.

Development of Airway Epithelium

Over 40 different cell types have been identified in the lungs of adult mammals. At least 8 of these cell types are found in the epithelium lining of the tracheobronchial conducting airways1,2 and include ciliated cells, basal cells, mucous goblet cells, serous cells, Clara cells, small mucous granule cells, brush cells, neuroendocrine cells, and a variety of partially differentiated cell types. The abundance and distribution of these cells varies throughout the conducting airways and differs by species.

Cytodifferentiation of two epithelial populations of the respiratory bronchiole during fetal lung development in the rhesus monkey.

This study describes the cytodifferentiation of the two populations of epithelial cells found in the respiratory bronchiole of the adult rhesus monkey. One population, pseudostratified and containing ciliated, nonciliated secretory, and basal cells, is found overlying the pulmonary artery (PA). The other population, not associated with the PA, contains nonciliated cuboidal cells between alveolar outpockets. In this study we used terminal conducting airways from the lungs of fetal (90 to 155 days gestational age [DGA]), postnatal, and adult rhesus monkeys. Ciliated cells were partially differentiated at 90 DGA (54% gestation) and completely differentiated by 134 DGA (80% gestation). Nonciliated secretory cells were partially differentiated at 95 DGA (57% gestation) but did not lose all glycogen until the postnatal period. Basal cells appeared by 134 DGA (80% gestation) and matured in the postnatal period. Small mucous granule cells appeared at 125 DGA (74% gestation) and did not change throughout fetal development. Neuroendocrine cells were present throughout the entire period studied. Nonciliated cuboidal bronchiolar cells of the nonciliated population of the respiratory bronchiole appeared at 105 DGA (62% gestation) and matured in the postnatal period. We conclude that 1) although most of the differentiation of the lower airway occurs before birth, most of the cell types are not completely differentiated at birth; 2) the sequence of differentiation for the cells of the ciliated pseudostratified epithelial population is ciliated, nonciliated secretory, and basal; 3) the sequence of differentiation for the nonciliated secretory cell is similar to that of the secretory cells in more proximal airways; and 4) basal, neuroendocrine, and small mucous granule cells are not a part of the differentiation sequence of the other cell types.

Tracheobronchial epithelium in the adult rhesus monkey: a quantitative histochemical and ultrastructural study.

Previous studies of the intrapulmonary conducting airways of sheep and rabbit have demonstrated marked diversity in the epithelial populations lining them. Because studies of trachea and centriacinar regions of macaque monkeys suggested that primates may be even more diverse, the present study was designed to characterize the epithelial population throughout the airway tree of one primate species, the rhesus monkey. Trachea and intrapulmonary airways of the right cranial and middle lobes of glutaraldehyde/paraformaldehyde-infused lungs of five adult rhesus monkeys were microdissected following the axial pathway. Each branch was assigned a binary number indicating its specific location within the tree. The trachea and six generations of intrapulmonary airway from the right cranial lobe were evaluated for ultrastructure and quantitative histology as were those of the right middle lobe for quantitative carbohydrate histochemistry. Four cell types were identified throughout the tree: ciliated, mucous goblet, small mucous granule, and basal. The tallest epithelium lined the trachea; the shortest, the respiratory bronchiole. The most cells per unit length of basement membrane were in proximal intrapulmonary bronchi; the least, in the respiratory bronchiole. The nonciliated bronchiolar epithelial or Clara cell was restricted to respiratory bronchioles. Sulfomucins were present in the vast majority of surface goblet cells in the trachea and proximal bronchi. In proximal bronchi, neutral glycoconjugates predominated in glands and acidic glycoconjugates in surface epithelium. In terminal and respiratory bronchioles the ratio of acidic glycoconjugate to neutral glycoconjugate equaled that in proximal bronchi, although glands were not present. Sulfomucins were minimal in terminal airways. We conclude that the characteristics of the epithelial lining of the mammalian tracheobronchial airway tree are very species-specific. The lining of the rhesus monkey does not have the diversity in cell types in different airway generations observed in sheep and rabbit. Also, the populations lining these airways in the rhesus are very different from either the sheep or rabbit in number, proportions of different cell types, glycoconjugate content, and distribution of specific cell types.

Effect of cigarette smoke condensate and vitamin A depletion on keratin expression patterns in cultured hamster tracheal epithelium

Keratin expression in hamster tracheal epithelium was investigated during organ culture in serum-free, hormone-supplemented medium using monospecific monoclonal antibodies. Generally, tracheal basal cells expressed keratins detected by antibodies RCK102 and RCK103, while columnar epithelial cells were stained positively by RGE53, RCK103, RCK105 and HCK19. Metaplastic squamous cell foci reacted with antibodies RKSE60, RCK103 and HCK19. Early metaplastic alterations were more clearly RKSE60-positive than the mature lesions. In the vitamin A-depleted tracheas basal cells were clearly RCK102-positive. Superficial cells in the central part of areas of squamous metaplasia induced by cigarette smoke condensate expressed the basal cell keratins, and were negative for the columnar cell keratin 18 detected by the RGE53 antibody. This finding suggests that in cigarette smoke condensate-induced squamous metaplasia basal cells play an important role. The mucus-producing cells at the edges of metaplastic squamous cell foci expressed the keratins specific to columnar cells. Cigarette smoke condensate exposure accelerated epithelial keratinization compared to the vitamin A-depleted epithelium. It was concluded that not only small mucous granule cells, but also basal cells are involved in the development and maintenance of induced squamous metaplasia in tracheal epithelium. Furthermore, in vitro vitamin A-depleted epithelium did not coexpress vimentin in addition to the different keratins.

Ciliated cells in vitamin A-deprived cultured hamster tracheal epithelium do divide.

The pseudostratified tracheal epithelium, composed of a heterogeneous phenotypically varying cell population, was studied with respect to the in vitro cell proliferative activity of differentiated epithelial cells. Ciliated tracheal epithelial cells so far have been considered to be terminally differentiated, nonproliferating cells. Tracheal organ cultures obtained from vitamin A-deprived Syrian Golden hamsters were cultured in a vitamin A-deficient, serum-free, hormone-supplemented medium. In vitamin A-deprived tracheal epithelium treated with physiologically active all-trans retinol and low cigarette-smoke condensate concentrations it is possible to stimulate the cell proliferation of both basal and columnar cells. Therefore, the probability of finding proliferating columnar cells was increased compared with the in vivo and the vitamin A-deprived situation in which cell proliferative activity is relatively low. In the presence of cigarette-smoke condensate in a noncytotoxic concentration, basal, small mucous granule, ciliated, and indifferent tracheal epithelial cells incorporated [methyl-3H]-thymidine into the DNA during the S phase. The finding that ciliated cells were labeled was supported by serial sections showing the same labeled ciliated cell in two section planes separated by 2 to 3 micron, without labeled epithelial cells next to the ciliated cell. Furthermore, a ciliated tracheal epithelial cell incorporating [methyl-3H]thymidine into DNA was also seen in tracheal cultures of vitamin A-deprived hamsters treated with all-trans retinol in a physiologic concentration.

Immunoelectron microscopic study on the histogenesis of epidermoid metaplasia in respiratory epithelium.

To elucidate the histogenesis of epidermoid metaplasia with an immunoelectron microscopic method, we studied the localization of secretory component (SC), a differentiation marker of the epithelial cell, in human bronchial epithelium. In the area of normal epithelium, SC was synthesized by mucous cells, particularly by small mucous granule cells. It was also found with small mucous granules in both superficial columnar or cuboidal cells in the area of stratification and superficial, less flattened cells in the area of epidermoid metaplasia, but SC was not identified in polygonal cells under the superficial SC-positive cells in both of the above areas. These superficial cells containing SC and small mucous granules possessed increasing number of tonofilaments parallel with the degree of flattening of the surface cells. On the other hand, in the advanced epidermoid metaplasia, in which surface cells were composed of very flattened cells, SC and small mucous granules were not found in the bronchial epithelium. Tonofilaments increased in number in the cytoplasm parallel to morphologic changes from basal cells to outermost flattened cells, via polygonal cells. These findings suggest that small mucous granule cells containing SC could gradually change to flattened cells through a quantitative change of a cytoplasmic component, such as SC or tonofilaments, and constitute the superficial cells of initial epidermoid metaplasia, in which surface cells are composed of less flattened cells. Moreover, these findings suggested that basal cells play an important role in epidermoid differentiation.

Correlated histochemical and electron microscopic studies of the esophageal epithelium in the salamander, Hynobius nebulosus.

The esophageal epithelium of the adult salamander, Hynobius nebulosus, was studied by histochemistry, including periodic acid-Shiff (PAS), alcian blue both at pH 2.5 and pH 1.0, and alcian blue (pH 2.5)-PAS with or without neuraminidase-digestion, and scanning and transmission electron microscopy. The epithelium was columnar in type, comprising partly pseudostratified and partly two-layered architecture; it consisted mainly of ciliated cells, goblet cells, and basal cells. The ciliated cells consisted of two types, light and dark: both types, especially the latter, frequently contained numerous small mucous granules in their apical portion. Ciliated-mucous cells were also occasionally present. Immature ciliated cells were frequently found. Goblet cells were divided into two types: Type I cells possessed electron lucent mucous granules, which frequently contained dense specific inclusions, and frequently bulged into the lumen; Type II cells had moderately electron-dense mucous granules with no inclusions and a conical apex which did not exceed the level of the lumen. The Type I cells were closely distributed throughout the esophagus, while the Type II were mainly dispersed in the cranial portion, remarkably decreasing in the caudal portion. Correlated histochemical and electron microscopic observations suggested that, in the Type I cell, mucous granules contain acid mucosubstances, while in the Type II, they possess neutral mucosubstances, and that in the Type I cell mucous granules consist of sialic acid-containing glycoproteins and their swollen portions are more highly sulfated than the non-swollen ones.

The tracheobronchial epithelium of the bonnet monkey (Macaca radiata): a quantitative ultrastructural study.

Since there are major differences between the airway epithelium of man and that of common laboratory species, the tracheobronchial epithelium of the bonnet macaque was characterized to evaluate its usefulness as a model for study of human conducting airways. This study compared the light microscopic, scanning electron microscopic, and ultrastructural appearance of epithelium from the posterior membranous and anterior cartilaginous trachea and mainstem bronchus. Population densities, epithelial volumetric densities, and frequency distributions of cross-sectional areas of nuclei were determined for cell types present on electron micrographs. Four epithelial cell types were distinguished by ultrastructural criteria. Basal cells were 31% of the population and were similar to those described in other species. Ciliated cells were also similar to those of other species and composed 41% of the population; their nuclei were larger than those of other cell types. Mucous goblet cells had large numbers of secretory granules with electron-dense cores and a lucent periphery. They were only 8% of the population by nuclear count but composed 20% of the epithelial volume. The fourth cell type had multiple small vesicles containing small amounts of granular material and was termed a "small mucous granule cell." Small mucous granule cells (16% of the population) were present in greater numbers than mucous goblet cells but were a smaller proportion of the epithelial volume (8%). While population densities of cell types determined from transmission electron micrographs did not vary between sample sites, scanning electron microscopy demonstrated longitudinal streaks of secretory cells in the posterior trachea suggesting that regional differences in epithelial organization exist. We conclude that the macaque extrapulmonary airway epithelium differs from published descriptions of laboratory rodents in both cell types present and relative abundance of those cell types. Although detailed quantitative studies of human extrapulmonary airways are not available, the primate airways resemble those of man in both the types of cells present and the complexity of pseudostratification.

Colon Epithelium. I. Light Microscopic, Histochemical, and Ultrastructural Features of Normal Colon Epithelium of Male Fischer 344 Rats<xref ref-type="fn" rid="fn2">2</xref><xref ref-type="fn" rid="fn3">3</xref><xref ref-type="fn" rid="fn4">4</xref>

Although the colon of the inbred F344 rat is not distinctly demarcated into ascending, transverse, and descending segments as in the human colon, it can roughly be divided into ascending and descending portions that show distinct light microscopic, histochemical, and ultrastructural features. The ascending colon is characterized by a "herringbone" pattern of mucosal folds and test-tube-shaped uniform crypts that contain mucous cells (MC) with abundant mucin (acidic mucopolysaccharide--mostly sialomucin) in the lower one-third of the crypt, whereas the upper one-third contains two putative cell populations: 1) MC containing large globules of neutral mucopolysaccharide or sulfomucin and 2) columnar cells (CC), the full capabilities of which are unknown. The descending colon has longitudinal folds and contains sparse MC with small mucous granules at the lower one-third of the crypt, whereas the upper one-third contains numerous goblet cells. Neutral mucopolysaccharide is sparse and the acidic mucin is exclusively sulfated. Histochemically, the descending segment of the rat colon resembles the human descending colon in that the predominant type of mucus is sulfomucin. Ultrastructurally, the cell types observed in both the ascending colon and the descending colon are: a) MC, b) CC, c) endocrine cells, and d) undifferentiated cells.

Chapter 1 Methods of Organ Culture for Human Bronchus

Publisher Summary This chapter discusses the methods of organ culture for human bronchus. One goal of organ culture is the maintenance of in vivo characteristics and specific cellular relationships. Both the human and bovine trachea and bronchus were cultured as 1-cm-square pieces in 60-mm Falcon plastic petri dishes with the epithelial surface uppermost. The culture medium was added to just reach the epithelial surface of the explant. Normal human bronchial epithelium is composed of a single layer of pseudostratified cells. The tall, columnar ciliated cells are the most numerous. Tall, columnar mucous cells are of two types: (1) goblet cells whose apices are dis tended with large mucous granules and (2) small mucous granule cells that have tiny mucous granules in the apex and are invisible by light microscopy unless stains for mucosubstances are used. The xeno-transplant studies show that the culture-induced metaplastic changes are reversible, because when such CMRL-cultured explants are transplanted into nude mice, the epithelium reverts to normal and columnar differentiation takes place and is maintained for over 1 year.

The Respiratory Epithelium. I. Human Bronchus<xref ref-type="fn" rid="FN2">2</xref><xref ref-type="fn" rid="FN3">3</xref>

Six morphologic cell types comprise the human bronchial epithelium: basal cells that do not reach the bronchial lumen, neurosecretory cells (Kulchitsky's cells, K-cells, or small granule cells) that rarely reach the lumen, and indifferent cells, mucous cells [small mucous granule cells (SMGC) and mucous goblet cells], ciliated cells, and ciliated-mucous cells that do reach the lumen. Ciliated-mucous cells bearing fully developed cilia and containing mucous granules are seen only occasionally. Three of the cell types that reach the lumen are microvillus covered and do not bear cilia. The microvillus-covered nonciliated cells are: 1) neurosecretory cells, 2) indifferent cells, and 3) mucous cells. Neurosecretory cells contain characteristic dense core granules. Such cells rarely reach the lumen. Indifferent cells are rarely seen. They have a pale cytoplasm and show no evidence of either ciliary or mucous differentiation. Similar cells are observed showing early signs of either ciliary or mucous differentiation or even both types of differentiation in the same cell. Mucous cells comprise the vast majority of microvillus-covered cells. They present either as SMGC with a few small mucous granules or as goblet cells, filled with mucus. These columnar cells are characterized ultrastructurally by dense cytoplasm and a well-developed endoplasmic reticulum and Golgi apparatus. The microvilli are coated with a glycocalyx that binds colloidal iron more avidly than that of either cilia or microvilli of ciliated cells. Possible interrelationships between the different cell types in normal epithelium are discussed.

The Respiratory Epithelium. II. Hamster Trachea, Bronchus, and Bronchioles<xref ref-type="fn" rid="FN2">2</xref><xref ref-type="fn" rid="FN3">3</xref>

The normal female hamster respiratory epithelium at five airway levels was characterized with the use of coordinated morphologic and histochemical techniques. Five morphologic cell types were recognized in the trachea, stem bronchi, and primary bronchl: basal cells and neurosecretory cells that were basally located and did not reach the lumen and mucous cells [mucous goblet cells and small mucous granule cells (SMGC)], indifferent cells showing mucous-ciliary differentiation, and ciliated cells that reached the lumen. Two epithelial cell types were observed in the bronchioles, ciliated cells and nonciliated Clara cells, both of which reached the lumen. Mucous cells presented as either SMGC with a few small periodic acid-Schiff-positive granules (diastase-resistant neutral mucosubstances) or as goblet cells, filled with the same material. Mucous cells were columnar, and the cytoplasm was electron-dense and contained a well-developed endoplasmic reticulum and Golgi complex. The microvilli of the mucous cells were coated more thickly with colloidal iron than either the cilia or microvilli of ciliated cells. Approximately one-half the cells in the trachea, bronchi, and bronchioles were ciliated. Ciliated cells containing intracellular ciliated cysts with normal cilia projecting into a closed space or ciliated cells bearing compound cilia were observed infrequently. Neurosecretory cells were rarely observed. These cells contained characteristic dense-core granules.

The cells of the rat gastric groove and cardia. An ultrastructural and carbohydrate histochemical study, with special reference to the fibrillovesicular cells.

The distal wall of the groove between the rat forestomach and glandular stomach is lined with a special type of columnar cells (CCGG) and with fibrillovesicular cells (FVC). The cardiac glands contain cardiac mucosa (CMC) and serous cells (CSC). The CCGG contain small mucous granules and special vesicles and tubules. The CMC are filled with large mucous granules and resemble mucous neck cells. The CSC are filled with large proteinaceous granules. The FVC are characterized by long microvilli, apical bundles of microfilaments and a complex "tubulovesicular system". The pattern of 3H-thymidine incorporation and the presence of immature and transitional forms indicate a possible origin of all the cell types concerned from a common undifferentiated precursor. The membranes of the tubulovesicular system of FVC as well as the apical cell membrane were reactive to Thiéry's carbohydrate stain. However, lanthanum tracing of the extracellular space and ultrastructural stereoscopy did not reveal a permanent continuity between both membrane systems. The absence of 3H-thymidine label showed that FVC were not proliferative. The structural characteristics of FVC do not account for a secretory, resorptive or receptive function. The special arrangement of microfilaments and the tubulovesicular system suggests an ability to fast changes in surface area.

Horny cell formation in the epidermis of Rana pipiens.

AbstractWell preserved transitional cells were found between differentiated cells and horny cells of the frog epidermis, thus facilitating the study of the sequential events involved in horny cell formation. Autolysosomes appear to play an important role in the formation of horny cells. These structures preferentially digest those cytoplasmic components which are not necessary constituents of the terminal horny cell. The release of the contents of the small mucous granules into the intercellular spaces is one of the initial events in horny cell formation. Filaments and large mucous granules seem to be resistant to the lytic digestion and contribute to the bulk of the horny cell. Loss of fluids through the plasma membrane and consolidation of the remaining constituents, results in a flattened horny cell. The appearance of a thickened membrane around the horny cell signifies the completion of the transformation process.